Cited by 155PI3K/AKT/mTOR signaling in cancerSignaling Pathways in DiseaseProtein Kinase Regulation and GTPase Signaling

The FKBP12-Rapamycin-binding Domain Is Required for FKBP12-Rapamycin-associated Protein Kinase Activity and G1 Progression

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Vilella-Bach, Montserrat, Nuzzi, Paul, Fang, Yimin, Chen, Jie·1999·Journal of Biological Chemistry

Abstract

The immunosuppressant rapamycin, in complex with its cellular receptor FKBP12, targets the cellular protein FKBP12-rapamycin-associated protein/mammalian target of rapamycin/rapamycin and FKBP12 target 1 (FRAP/mTOR/RAFT1) and inhibits/delays G1 cell cycle progression in mammalian cells. As a member of the novel phosphatidylinositol kinase-related kinase family, FRAP's kinase activity is essential for its signaling function. The FKBP12-rapamycin binding (FRB) domain in FRAP is also speculated to play an important role in FRAP function and signaling. However, the biochemical and physiological functions of FRB, as well as the mechanism for rapamycin inhibition, have been unclear. The present study focuses on investigation of FRB's role and the functional relationship between FRB domain and kinase domain in FRAP. Microinjection of purified FRB protein into human osteosarcoma MG63 cells results in a drastic blockage of the G1 to S cell cycle progression; such a dominant negative effect is reversed by a point mutation (Trp2027 --> Phe). The same mutation also abolishes kinase activity of FRAP without affecting ATP binding, and truncation studies suggest that upstream sequences including FRB are required for kinase activity in vitro. Given these data, we propose a model for FRAP function, in which the FRB domain is required for activation of the kinase domain, possibly through the interaction with an upstream activator. In addition, our observations provide direct evidence linking FRAP function to G1 cell cycle progression.

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Crossref · 10.1074/jbc.274.7.4266

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  • The mTOR kinase N-lobe features a large insertion corresponding to the FKBP12-rapamycin binding (FRB) domain specific to mTORC1 among the PIKKs.

    “Notably, however, the kinase N-lobe features a large insertion corresponding to the FKBP12-rapamycin binding (FRB) domain (Chen et al., 1995; Chiu et al., 1994; Vilella-Bach et al., 1999) specific to mTORC1 among the PIKKs.”

    From Signal integration in the (m)TORC1 growth pathway (2018) · cites this paper as Vilella-Bach et al., 1999

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Authors
Vilella-Bach, M., Nuzzi, P., Fang, Y., & Chen, J.
Affiliations
University of Illinois Urbana-Champaign (US)
Corresponding author
Chen, Jie
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1999-02
Volume
274
Issue
7
Pages
4266-4272
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2000281583
Cited by
154
References
48
Citations by year
2026: 2 · 2025: 5 · 2024: 3 · 2023: 6 · 2022: 7 · 2021: 7 · 2020: 6 · 2019: 5 · 2018: 3 · 2017: 4 · 2016: 3 · 2015: 8 · 2014: 3 · 2013: 9 · 2012: 3
FWCI
3.31
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › PI3K/AKT/mTOR signaling in cancer
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Signaling Pathways in Disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords
FRAP, mTOR, G1 cell cycle progression, rapamycin, immunosuppressants, dominant negative effect, osteosarcoma cells, ATP binding
MeSH terms
Amino Acid Sequence, Binding Sites, Carrier Proteins, Catalysis, Cell Line, Humans, Immunosuppressive Agents, Microinjections, Molecular Sequence Data, Protein Kinases, Serine, Tryptophan, Tumor Cells, Cultured, G1 Phase, S Phase, Mutagenesis, Site-Directed, Sequence Homology, Amino Acid, Phosphotransferases (Alcohol Group Acceptor), Phosphatidylinositol 3-Kinases, Amino Acid Substitution, Immunophilins, Sirolimus, Tacrolimus Binding Proteins, TOR Serine-Threonine Kinases

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